Particulate poly(glycidyl methacrylate-co-ethylene dimethacrylate) material for protein separation by anion-exchange chromatography

New materials with different structures are being developed in order to be used in protein separation by ion exchange chromatography. Materials for beds are becoming of smaller particulate size, at the expense of lower permeability, in order to obtain better peak resolution during separation. On the other hand, polymeric organic monoliths, obtained by “in situ” polymerization, have been synthethized in recent years for similar purposes. In this work, a variety of poly(glycidyl methacrylate-co-ethylene dimethacrylate) resin has been synthetised by “in situ” polymerization, milled and classified in order to verify its behaviour when used at particulate state in protein separation by ion exchange chromatography, even at semipreparative scale, which is not possible to carry out using monoliths. The particulate and classified material was aminofunctionalized with diethylamine and then properly characterized in order to determine its physical properties and evaluate its separation performances once packed in chromatographic columns. The results show that the material obtained presents good permeability and favourable equilibrium isotherms. Kinetics follows the pore diffusion model, giving diffusion coefficients similar to others reported for several commercial materials. The axial dispersion coefficients obtained at different flow rate allow to forecast a reasonably good behavior in the anion exchange chromatographic fractionation of proteins, as it was proved with egg white proteins at laboratory and semipreparative scale.     

A hydrophilic matrix for boron isolation: Monodisperse-porous poly(glycidyl methacrylate-co-ethylene dimethacrylate) particles carrying diol functionality

The novel chelating adsorbent has been synthesized by functionalization of monodisperse-porous poly(glycidyl methacrylate-co-ethylene dimethacrylate) particles with N-methylglucamine (NMDG) as a boron selective ligand. The resulting resin can adsorb boron in almost all pH range (pH 4–10) and its maximum capacity is ca. 12 mg/g. The adsorption behavior of poly(GMA-co-EDM)NMDG resin obeyed Langmuir isotherm well.     

Surface modification of macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) resins for improved Candida antarctica lipase B immobilization

Crosslinked macroporous hydrophilic poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [abbreviated poly(GMA-co-EGDMA)] with identical chemical structure (60% of glycidyl methacrylate) but with varied average pore sizes (from 30 to 560 nm), specific surface areas (from 13.2 to 106.0 m²/g), specific volumes (from 0.755 to 1.191 cm³/g) and particle sizes (<100 μm–630 μm) were synthesized via suspension polymerization. Modifications of poly(GMA-co-EGDMA) with various diamines (1,2-diaminoethane, 1,4-diaminobutane, 1,6-diaminohexane and 1,8-diaminooctane), 2-fluoroethylamine, glutaraldehyde and cyanuric chloride were carried out. The influence of the interaction between Candida antarctica lipase B (Cal-B) and various carriers during immobilization on the loading and hydrolytic activity (hydrolysis of para-nitrophenyl acetate) of the immobilized Cal-B were studied. Immobilization of Cal-B was performed at different temperatures and pH values. Cal-B immobilized at 30 °C and pH 6.8 was leading to increased activities. Purely physical adsorption between enzyme and copolymer was observed on carriers in which amine or fluorine groups were introduced into the carrier structure by modification with various diamines or 2-fluoroethylamine. As a consequence enzyme loading and activity decreases. In contrary, modification of the poly(GMA-co-EGDMA) with glutaraldehyde and cyanuric chloride results in a covalent connection between enzyme and carrier. The obtained results show a significant increase in Cal-B activity. The influence of the amount of glutaraldehyde and cyanuric chloride used for modification was screened. Increasing the amount of glutaraldehyde or cyanuric chloride used for modification resulted in an increase of the enzyme loading. Consequently, higher amount of glutaraldehyde used led to a higher fraction of the enzyme molecules that are covalently connected on to the carrier. As the amount of glutaraldehyde or cyanuric chloride used for modifications increases, activity of immobilized C. antarctica lipase B primarily increases, showing the highest value for 0.66% and 0.050% w/w, respectively, and subsequently decreases. We could show that Cal-B immobilized on epoxy-containing copolymer modified with glutaraldehyde and cyanuric chloride performs higher activity than free enzyme powder.     

Immobilization of mushroom polyphenol oxidase on poly(allyl glycidyl ether-co-ethylene glycol dimethacrylate) macroporous beaded copolymers

Functional, macroporous, beaded copolymers containing epoxy groups were synthesized for immobilization of polyphenol oxidase (PPO) from edible mushroom (Agaricus bisporus). The effect of incorporation of two different sets of monomers such as glycidyl methacrylate (GMA) and allyl glycidyl ether (AGE) and the effect of cross-linking agent ethylene glycol dimethacrylate (EGDM) with varying cross-link densities on binding and expression of mushroom PPO activity were studied. The effect of porogen viz. cyclohexanol and hexanol on PPO immobilization was studied. AGE copolymers with hexanol as a porogen were found to give higher binding and expression of PPO activity than GE polymers. Cross-linking of amino groups of enzyme with 5% glutaraldehyde for 6 h gave a stable binding of PPO on AGE-75(Hex) polymer with storage half-life of approximately 25 days. Under optimum conditions, AGE-75(Hex) polymer gave 70.3% of activity yield while percent retention of PPO activity was found to be 83.5%. Immobilized PPO showed a broader pH, higher temperature and excellent storage stability.     

Reactive polymers, 47. Sorption properties of the reaction product of macroporous poly (glycidyl methacrylate-co-ethylene dimethacrylate) with 8-aminoquinoline

The sorption of Fe(III), Al(III), Mn(II), Co(II), Ni(II), Cu(II), Zn(II), Au(III), Pd(II), Pt(IV), and Hg(II) ions on a sorbent prepared by reacting the copolymer of glycidyl methacrylate and ethylene dimethacrylate with 8-aminoquinoline was investigated. Gold and palladium are strongly absorbed in hydrochloric acid. Results of static and dynamic tests indicate the possibility of separation of Au(III) and Pd(II) from Pt(IV) and from the other metal ions investigated in the study. The sorption of Hg(II) by the polymer increases in nitric acid.     

Reactive polymers,XXXVII. An investigation of the internal structure of polymeric sorbents based on poly(2,3-epoxypropyl methacrylate-co-ethylene dimethacrylate)

The internal structure of polymeric sorbents based on 2,3-epoxypropyl methacrylateethylene dimethacrylate was investigated by means of mercury porosimetry and by employing the sorption method. The results were confronted with characteristic chromatographic data and helped to elucidate the relationships existing between the internal structure of copolymers and their chromatographic properties.     

Oligoamine-functionalised poly(glycidyl methacrylate-ethyleneglycol dimethacrylate) resins as moderate base extractants for gold from cyanide solutions

A series of oligoamines have been attached to poly(glycidyl methacrylate) resins to yield a corresponding group of moderate base resins. Species carrying ethylenediamine, diethylenetriamine, triethylenetetramine, N,N′-bis(3-aminopropyl)ethylenediamine, and 3,3′-aminobispropylamine and phenyl-2-imidazoline are described. These have been examined as extractants for gold from alkaline cyanide solutions, along with a commercially available polystyrene-based resin carrying a 2,5,7-triazabicyclo[4,4,0]dec-5-ene functionality. Feed solutions containing only gold cyanide and also gold in the presence of the base metals Cu, Co and Ni, have been examined. The stripping of metal loaded resins using various concentrations of NaOH has also been investigated. The results of loading and stripping experiments are discussed in the light of the basicity of the various oligoamines, and also in the context of relevant data already available in the literature. Overall, though very effective gold extraction has been demonstrated with high selectivity relative to base metals, efficient stripping is problematical and has been demonstrated in only a few cases. This suggests that gold extraction and stripping does not involve only protonation/deprotonation of the moderate base functions, but extraction probably involves a contribution from gold reduction as well.     

Immobilization of Myceliophthora thermophila laccase on poly(glycidyl methacrylate) microspheres enhances the degradation of azinphos-methyl

Laccase enzymes are multicopper oxidases capable of oxidizing different compounds. However, to be able to use this biocatalyst for industrial applications, their immobilization is needed. The present work investigates the immobilization of Myceliophthora thermophila laccase (MtL) on monodisperse microspheres of poly(glycidyl methacrylate) (PGMA) to be used in azinphos-methyl degradation. The immobilization was optimized to achieve the highest activity of the immobilized enzyme. As result, the protein load obtained was 2.5 mg protein g⁻¹ carrier (35 U g⁻¹ of carrier). The immobilized enzymes showed a broadened pH and temperature range of optimum activity and significantly improved the storage and operational stability. Finally, the complete degradation of azinphos-methyl using immobilized laccase was achieved after 1 h of reaction. The collected data indicate that the immobilization of MtL on PGMA microspheres is an excellent alternative to improve biochemical properties in the enzyme and to allow their efficient use in pesticides degradation. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47417.     

Stable poly(glycidyl methacrylate‐co‐ethylene glycol dimethacrylate) microspheres via precipitation polymerization

Narrow‐dispersion or monodisperse with stable and smooth surface polymer microspheres were prepared without a significant coagulum by precipitation polymerization in the absence of any stabilizer. The monomer glycidyl metharylate (GMA) was copolymerized with ethyleneglycol dimethacrylate (EGDMA) as crosslinker by precipitation polymerization technique with 2,2′‐azobisisobutyronitrile as initiator in neat acetonitrile. The effects of the content of EGDMA on the polymerization characteristics and size/uniformity of the microspheres were investigated. The onset of the thermal degradation temperature at higher temperature and the swelling test suggest that the prepared particles were highly crosslinked. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Microwave-assisted synthesis of poly(glycidyl azide-co-tetrahydrofuran)

The poly(glycidylazide-co-terahydrofuran) [poly(GA-THF)] or GA-THF copolymers are recognized and established as high-energy binders in explosive systems. In present study, we have developed a greener approach for the syntheses of poly(glycidylazide-co-tetrahydrofuran) copolymers using microwave (MW) irradiation technique. Polymerization of epichlorohydrin and tetrahydrofuran was carried out in the presence of borontrifluoride etherate and ethylene glycol to afford polyepichlorohydrin-tetrahydrofuran (PECH-THF) copolymer followed by its reaction with sodium azide in DMF under microwave irradiation to give poly(GA-THF). The poly(GA-THF) copolymer and their precursors were characterized by various spectral techniques such as MS, ¹H-NMR, ¹³C-NMR, UV–visible, and FTIR. The thermal analysis was done by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Molecular weights of these copolymers were determined using GPC techniques. Morphology of these copolymers was visualized using SEM. Our experimental study shows that the microwave irradiation was effective for the second step (azidation reaction) than conventional heating for the synthesis of GA-THF copolymers. Also, the benign syntheses were tackled and explored with range of solvents. Thus, the current methodology, might advent the researchers dealing with these high-energy binder materials in future.

     

聚缩水甘油醚硝酸酯研究进展

综述了一种在新一代高能航天固体推进剂、战术发动机高能低特征信号推进剂和聚合物黏结炸药(PBX)等领域中备受关注的含能聚合物黏合剂——聚缩水甘油醚硝酸酯(PGN)黏合剂的合成、性能及应用研究进展。     

复合纤维素固定化脲酶的研制

将脉酶吸附在纤维素载体上，再经过交联和包埋双重固定技术制备出活性高和稳定性好的复合纤维素固定化脲酶。     

Preparation of amidoximated poly(glycidyl methacrylate) microbeads

Poly(glycidyl methacrylate) (PGMA) microbeads were synthesized by a simple suspension polymerization of glycidyl methacrylate and ethylene glycol dimethacrylate. The epoxy groups of the microbeads were firstly modified with 3,3′‐iminodipropionitrile (IDPN) and the resulting nitrile groups were then converted to amidoxime. From scanning electron microscopy studies, the average size of the PGMA microbeads was determined as 170 µm, which was not changed by the modification processes. For the modification of epoxy groups with IDPN, the intensity of the C≡N absorption band at 2249 cm^?1 increased proportionally with reaction time; for the conversion of nitrile groups to amidoxime, it decreased. Attenuated total reflectance Fourier transform infrared spectroscopy measurements clearly showed the disappearance of the original nitrile groups and the formation of amidoxime groups through treatment with hydroxylamine under the reaction conditions specified. The microbeads possessed good thermal and morphological properties and chemical stability suitable for practical use. Therefore, the amidoximated PGMA microbeads could be used in batch and continuous processes for the adsorption of uranyl ions from seawater or aqueous media. Copyright © 2010 Society of Chemical Industry     

Immobilization of penicillin acylase on copolymer of butyl acrylate and ethylene glycol dimethacrylate

The effects of glutaraldehyde, enzyme concentrations and reactants volumes, ionic strength, pH value and carrier particle diameter on immobilization of penicillin acylase onto acrylic carriers were studied. The activity of immobilized enzyme preparations was also studied over a range of pH values and temperatures and thermal and pH stabilities were determined. The use of the immobilized preparation for penicillin G hydrolysis in a batch reactor was investigated. The immobilized enzyme gave a significant reduction in hydrolysis time compared to hydrolysis by the native enzyme.     

Immobilization of polymers on cellulose acetate membranes

Using models of dead-end filtration theory, the kinetics of forming dynamic layers of sulfate lignin (SL) and sodium carboxy-methylcellulose (Na-CMC) onto CA membranes during cross-flow filtration of dilute solutions of polymers was studied. It was found for both polymers (compact SL and linear Na-CMC), that the polymer layer with the least hydraulic resistance, which yields a small reduction in membrane water permeability (10–20%), but a significant increase in salt rejection, is formed, if the process kinetics corresponds to J-V linear dependence predicted by the model of 'blocking a pore by a single particle's. The results obtained may be used to define the optimum conditions for immobilizing the available catalytic active polymers on regular semipermeable membranes during the membrane filtration process.     

聚甲基丙烯酸缩水甘油酯疏水纳凝胶的制备与表征

采用乳液聚合法制备了聚甲基丙烯酸缩水甘油酯疏水纳凝胶。通过透射电子显微镜、傅里叶红外光谱和动态激光光散射等测试对其结构和形貌进行了表征,对疏水纳凝胶形成过程规律和对温度、pH和时间稳定性进行了考察,并探究了单体浓度、交联剂含量和乳化剂浓度对纳凝胶粒径的影响规律。结果表明：所得疏水纳凝胶具有粒径均一、分散稳定和溶胀性能好等优点。疏水纳凝胶粒径在一定温度和pH范围内无变化,在30天内粒径从76nm增至116nm,zeta电位随温度升高而增加,其值稳定在-90~-30mV之间,说明是一种较为稳定的凝胶体系。在实验范围内,纳凝胶粒径随单体甲基丙烯酸缩水甘油酯和交联剂二甲基丙烯酸乙二醇酯浓度的增加从80nm增至250nm,随乳化剂浓度增加从230nm降至60nm。     

Degradable poly(ester amine) based on poly(ethylene glycol) dimethacrylate and polyethylenimine as a gene carrier

Degradable poly(ester amine) (PEA) based on poly(ethylene glycol) dimethacrylate (PMEG) and polyethylenimine (PEI) were synthesized by Michael addition reaction. The ratios of PEI to PMEG in PEAs were 0.99, 1.02, and 1.07 with corresponding number‐average molecular weight of 1.3 × 10⁴, 1.2 × 10⁴, and 0.9 × 10⁴, respectively. Degradation rate of PEA at pH 7.4 was higher than that at pH 5.6. Good plasmid condensation and protection ability was shown when N/P molar ratio of PEA to DNA was above 15 (N: nitrogen element in PEA, P: phosphate in DNA). PEA/DNA complexes had positive zeta potential, narrow size distribution, good dispersity, and spheric shape with size below 250 nm when N/P ratio was above 30, suggestion of their endocytosis potential. Compared with PEI 25 KDa, the PEAs showed essential nontoxic to HeLa, HepG2 and 293T cells. With an increase in the molecular weight of PMEG, the transfection efficiency of PEAs in HeLa, HepG2 and 293T showed a tendency to decrease as well as the percent decrease of gene transfection efficiency with serum. The mechanism of PEA‐mediated gene transfection was attributed to “proton sponge effect” of PEI in the PEA. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Composites of allyl glycidyl ether modified polyethylene and cellulose

Linear medium density polyethylene (LMDPE) was functionalized with allyl glycidyl ether (AGE) in an internal laboratory mixer in the presence of peroxide. AGE is a bifunctional monomer, which forms unstable and energetically rich macroradicals. Upon increasing the peroxide content chain scission and grafting yield were favored. The degree of functionalization was determined by means of a calibration function for Fourier-transformed infrared spectroscopy (FTIR). Grafting AGE onto LMDPE led to a small loss of crystallinity, as evidenced by differential scanning calorimetry (DSC) and X-ray diffractometry analyses. Composites of LMDPE or functionalized LMDPE-AGE and cellulose were prepared in the mixer with filler contents ranging from 20 to 50 wt%. Composites of AGE functionalized LMDPE and filler content higher than 30 wt% presented tensile properties superior to that observed for composites with unmodified LMDPE. Scanning electron microscopy (SEM) on the composites fracture surface evidenced good interfacial adhesion between LMDPE-AGE and cellulose fibers.     

Hydrogels from glycidyl derivatives of poly(vinyl alcohol)

Poly(vinyl alcohol) (PVA) was modified with phthalic anhydride to obtain half esters with carboxylic acid groups, which made the reaction with epichlorohydrin easier. The oxirane ring underwent a further crosslinking that led to crosslinked polymers with polar groups capable of interacting strongly with water and therefore with properties of hydrogels. The curing kinetics of the crosslinking were studied by differential scanning calorimetry, and the dependence of the activation energy on conversion degree was studied by isoconversional kinetic analysis. Water absorption was determined gravimetrically as a function of time at room temperature. The swelling behavior of these hydrogels was related to the degree of crosslinking. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 693–698, 2003     

Immobilization of glucoamylase on magnetic poly(styrene) particles

Magnetic poly(styrene) particles including active groups were prepared for enzyme immobilization without any activation process. Glucoamylase, which is widely used in industry, was immobilized onto these particles. The effects of pH, buffer concentration, and temperature on immobilization were investigated; moreover, the effect of immobilization temperature on immobilized glucoamylase activity was determined for the hydrolysis of maltose. The acetate buffer with the concentration of 6 × 10⁻⁴ M at pH 4 and 20–30°C was found as the most suitable medium for the immobilization of the glucoamylase. The amount of bound protein is 8 mg/g particle with the immobilization yield of 70%. The maximum activity obtained with immobilized glucoamylase is approximately 70% of the free one. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 72: 69–73, 1999